33 research outputs found

    De Novo Assembly of the Dirofilaria immitis Genome by Long-Read Nanopore-Based Sequencing Technology on an Adult Worm from a Canine Cardiopulmonary Dirofilariosis Case

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    Dirofilaria immitis is a zoonotic parasitic nematode that infects domestic and wild canids, among its vertebrate hosts. The genetic analysis of D. immitis nowadays transcends the need for genetic taxonomy of nematodes, such as the study of resistance to macrocyclic lactone. We expanded the use of long-read nanopore-based sequencing technology on nematodes by performing genomic de novo assembly of a D. immitis specimen retrieved from a canine cardiopulmonary dirofilariasis case using the ONT MinION platform, followed by the study of macrocyclic lactone resistance. The assembled genome of D. immitis consists of 110 contigs with an N50 of 3687191. The genome size is 87899012 and contains a total of 9741 proteins; 6 ribosomal RNAs, with three belonging to the small subunit (18S) and three to the large subunit (28S); and 73 tRNAs. Subsequent analysis of six loci previously characterized as being associated to macrocyclic lactone resistance selection pressure showed that four have a genotype associated with either some loss of efficacy or the resistance phenotype. Considering the zoonotic potential of D. immitis, the identification of a resistant parasite alerts for the overuse of macrocyclic lactone in the region, which poses a potential risk to both veterinary and human public health. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.Funding: This work was supported by national funds through the Fundação para a CiĂȘncia e Tec-nologia (FCT, Portuguese Foundation for Science and Technology) under projects UIDB/04750/2020, LA/P/0064/2020, UIDB/CVT/00772/2020, and LA/P/0059/2020

    Pervasive hybridization with local wild relatives in Western European grapevine varieties

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    Grapevine (Vitis vinifera L.) diversity richness results from a complex domestication history over multiple historical periods. Here, we used whole-genome resequencing to elucidate different aspects of its recent evolutionary history. Our results support a model in which a central domestication event in grapevine was followed by postdomestication hybridization with local wild genotypes, leading to the presence of an introgression signature in modern wine varieties across Western Europe. The strongest signal was associated with a subset of Iberian grapevine varieties showing large introgression tracts. We targeted this study group for further analysis, demonstrating how regions under selection in wild populations from the Iberian Peninsula were preferentially passed on to the cultivated varieties by gene flow. Examination of underlying genes suggests that environmental adaptation played a fundamental role in both the evolution of wild genotypes and the outcome of hybridization with cultivated varieties, supporting a case of adaptive introgression in grapevine.info:eu-repo/semantics/publishedVersio

    The diacylglycerol kinase α/Atypical PKC/ÎČ1 integrin pathway in SDF-1α mammary carcinoma invasiveness

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    Diacylglycerol kinase α (DGKα), by phosphorylating diacylglycerol into phosphatidic acid, provides a key signal driving cell migration and matrix invasion. We previously demonstrated that in epithelial cells activation of DGKα activity promotes cytoskeletal remodeling and matrix invasion by recruiting atypical PKC at ruffling sites and by promoting RCP-mediated recycling of α5ÎČ1 integrin to the tip of pseudopods. In here we investigate the signaling pathway by which DGKα mediates SDF-1α-induced matrix invasion of MDA-MB-231 invasive breast carcinoma cells. Indeed we showed that, following SDF-1α stimulation, DGKα is activated and localized at cell protrusion, thus promoting their elongation and mediating SDF-1α induced MMP-9 metalloproteinase secretion and matrix invasion. Phosphatidic acid generated by DGKα promotes localization at cell protrusions of atypical PKCs which play an essential role downstream of DGKα by promoting Rac-mediated protrusion elongation and localized recruitment of ÎČ1 integrin and MMP-9. We finally demonstrate that activation of DGKα, atypical PKCs signaling and ÎČ1 integrin are all essential for MDA-MB-231 invasiveness. These data indicates the existence of a SDF-1α induced DGKα - atypical PKC - ÎČ1 integrin signaling pathway, which is essential for matrix invasion of carcinoma cells

    Uncovering new indicators to predict stability, maturity and biodiversity of compost on an industrial scale

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    Currently, the metagenomic study of the composting process has gained great importance since it has allowed the identification of the existence of microorganisms that, until now, had not been isolated during the process by traditional techniques. However, it is still complex to determine which bioindicators could reveal the degree of maturity and stability of a particular compost. Thereby, the main objective of this work was to demonstrate the possible correlation between traditional parameters of maturity and stability of compost, with other indicators of biodiversity in products highly heterogeneous from composting processes on an industrial scale. The results demonstrated the enormous influence of the raw materials in characterizing the products obtained. Even so, important relationships were established between the Chao1 and Shannon indexes, and certain parameters related to the maturity, stability and toxicity of the samples, such as nitrification index, humification rate, phenolic content, germination index or oxygen consumptio

    Usefulness of a New Large Set of High Throughput EST-SNP Markers as a Tool for Olive Germplasm Collection Management

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    Germplasm collections are basic tools for conservation, characterization, and efficient use of olive genetic resources. The identification of the olive cultivars maintained in the collections is an important ongoing task which has been performed by both, morphological and molecular markers. In the present study, based on the sequencing results of previous genomic projects, a new set of 1,043 EST-SNP markers has been identified. In order to evaluate its discrimination capacity and utility in diversity studies, this set of markers was used in a representative number of accessions from 20 different olive growing countries and maintained at the World Olive Germplasm Collection of IFAPA Centre ‘Alameda del Obispo’ (Córdoba, Spain), one of the world’s largest olive germplasm bank. Thus, the cultivated material included: cultivars belonging to previously defined core collections by means of SSR markers and agronomical traits, well known homonymy cases, possible redundancies previously identified in the collection, and recently introduced accessions. Marker stability was tested in repeated analyses of a selected number of accessions, as well as in different trees and accessions belonging to the same cultivar. In addition, 15 genotypes from a cross ‘Picual’ × ‘Arbequina’ cultivars from the IFAPA olive breeding program and a set of 89 wild genotypes were also included in the study. Our results indicate that, despite their relatively wide variability, the new set of EST-SNPs displayed lower levels of genetic diversity than SSRs in the set of olive core collections tested. However, the EST-SNP markers displayed consistent and reliable results from different plant material sources and plant propagation events. The EST-SNPs revealed a clear cut off between inter- and intra-cultivar variation in olive. Besides, they were able to reliably discriminate among different accessions, to detect possible homonymy cases as well as efficiently ascertain the presence of redundant germplasm in the collection. Additionally, these markers were highly transferable to the wild genotypes. These results, together with the low genotyping error rates and the easy and fully automated procedure used to get the genotyping data, validate the new set of EST-SNPs as possible markers of choice for olive cultivar identification

    Abstracts from the Food Allergy and Anaphylaxis Meeting 2016

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    PlantFuncSSR: Integrating first and next generation transcriptomics for mining of SSR-functional domains markers

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    et al.Analysis of repetitive DNA sequence content and divergence among the repetitive functional classes is a well-accepted approach for estimation of inter- and intra-generic differences in plant genomes. Among these elements, microsatellites, or Simple Sequence Repeats (SSRs), have been widely demonstrated as powerful genetic markers for species and varieties discrimination. We present PlantFuncSSRs platform having more than 364 plant species with more than 2 million functional SSRs. They are provided with detailed annotations for easy functional browsing of SSRs and with information on primer pairs and associated functional domains. PlantFuncSSRs can be leveraged to identify functional-based genic variability among the species of interest, which might be of particular interest in developing functional markers in plants. This comprehensive on-line portal unifies mining of SSRs from first and next generation sequencing datasets, corresponding primer pairs and associated in-depth functional annotation such as gene ontology annotation, gene interactions and its identification from reference protein databases.Gaurav Sablok thanks Plant Functional Biology and Climate Change Cluster (C3), University of Technology Sydney, PO Box 123, Broadway, NSW 2007, Australia, for providing the computational facilities. An internal grant number to GS (2226018) supported this work. JAH and TYS were partially supported by High Impact Research Chancellory Grant UM.C/625/1/HIR/MOHE/SCI/19 from the University of Malaya.Peer reviewedPeer Reviewe

    Conductas de riesgo en los adolescentes

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    La adolescencia es una etapa de cambios que se produce entre la niñez y juventud. EstĂĄ marcada por un cambio en las expectativas, en relaciĂłn no solamente con las funciones sexuales y reproductivas, sino tambiĂ©n con el status social. En esta etapa se dan con mayor intensidad conductas de riesgo que son aquellas acciones realizadas por el individuo o comunidad, que pueden llevar a consecuencias nocivas. El objetivo de la revisiĂłn es describir diferentes criterios de las conductas de riesgo en adolescentes tanto en Cuba como en el mundo, asĂ­ como realizar un acercamiento en las comunidades cerradas. Para la bĂșsqueda bibliogrĂĄfica se revisaron libros de texto y revistas biomĂ©dicas nacionales e internacionales, asĂ­ como protocolos, guĂ­as y programas existentes en Cuba y en otros paĂ­ses sobre la atenciĂłn integral a los adolescentes. En la actualidad existe diversidad de criterios al abordar las conductas de riesgo de los adolescentes. Esto ha hecho difĂ­cil el abordaje integral de la salud de los adolescentes. En las comunidades cerradas no se ha estudiado a fondo las conductas de riesgo de los adolescentes con un enfoque social. El desconocimiento sobre estos temas constituye una de las causas fundamentales que conllevan a conductas de riesgos en los adolescentes, por lo que debe ser estudiado e implementarse polĂ­ticas y programas de prevenciĂłn para disminuir estos comportamientos y sus consecuencias
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